1. The Structures of DNA and The Structures of DNA and RNARNA
2. GenomeGenomess
3. DNA ReplicationDNA Replication
4. DNA Repair and DNA Repair and RecombinationRecombination
5. Transcription and Transcription and ProcessingProcessing
6. TranslationTranslation
7. Control of Gene ExpressionControl of Gene Expression
8. Manipulating DNA and RNA
Introduction to
Molecular Genetics
http://priede.bf.lu.lv/ Studiju materiāli / MolekularasBiologijas / Ievads MolGen / EN
http://priede.bf.lu.lv/ Studiju materiāli / MolekularasBiologijas / Ievads MolGen / EN
• Electrophoresis– separates molecules by charge, size and structure
• Hybridization– detects specific DNA or RNA sequences through
complementary base-pairing
• Restriction endonucleases– cleave specific double-stranded (ds) DNA sequences
• DNA cloning (genetic engineering)– combines DNA from different sources
• Reverse transcription– makes DNA (cDNA = copy DNA) from RNA
• Mutagenesis– introduces mutations into genes
• Transgenic animals– allow elucidation of gene functions
• PCR– amplifies specific DNA fragments
• Sequencing– determines the nucleotide sequence of nucleic acids
Some techniques for DNA & RNA analysis
ELECTROPHORESIS separates molecules by size (& structure)
Molecular Biology of the Gene, 5th Edition
HYBRIDIZATION detects specific DNA or RNA sequences
Life The Science of Biology, 7th Edition
The principle of hybridization method
GENETIC ENGINEERING combines DNA from different sources
RESTRICTION ENDONUCLEASES cleave specific DNA sequences
GENETIC ENGINEERING combines DNA from different sources
Restriction fragments with compatible sticky ends easily join (anneal)
1
2
3
4
33
Which of the four black sticky ends will anneal to the red sticky end?
GENETIC ENGINEERING combines DNA from different sources
Restriction fragments with compatible sticky ends easily join (anneal)
333
GENETIC ENGINEERING combines DNA from different sources
Vectors transport recombinant DNA into the host cells
A vector should have:
The ability to replicate independently
Restriction sites
Reporter gene(-s)
A relatively small size
Life The Science of Biology, 7th Edition
GENETIC ENGINEERING combines DNA from different sources
Cloning of a DNA fragment into a plasmid vector
GENETIC ENGINEERING combines DNA from different sources
Expression vectors allow a foreign gene to be expressed in a host cell
Life The Science of Biology, 7th Edition
Mutations can be introduced into a gene using MUTAGENESIS
normāls proteīns proteīns, kas satur vēlamo aminoskābes apmaiņu
Reminder on the requirements for DNA synthesis (replication)
Molecular Biology of the Gene, 5th Edition
Polymerase Chain Reaction (PCR) amplifies specific DNA fragments
Molecular Biology of the Cell, 4th Edition
DNA SEQUENCING determines the nucleotide sequence of DNA fragments
Sanger (1975) dideoxy (chain termination) method
Molecular Biology of the Gene, 5th Edition
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